EP1406865B1 - Succinic acid derivatives - Google Patents

Succinic acid derivatives Download PDF

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Publication number
EP1406865B1
EP1406865B1 EP02738174A EP02738174A EP1406865B1 EP 1406865 B1 EP1406865 B1 EP 1406865B1 EP 02738174 A EP02738174 A EP 02738174A EP 02738174 A EP02738174 A EP 02738174A EP 1406865 B1 EP1406865 B1 EP 1406865B1
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Prior art keywords
alkyl
cycloalkyl
group
aryl
substituted
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EP02738174A
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German (de)
French (fr)
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EP1406865A2 (en
Inventor
Thomas RÖLLE
Thomas Lehmann
Markus Albers
Gerhard Müller
Gerhard Hessler
Rüdiger Fischer
Masaomi Tajimi
Karl Ziegelbauer
Kevin Bacon
Haruki Hasegawa
Hiromi Okigami
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Bayer AG
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Bayer Healthcare AG
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C233/00—Carboxylic acid amides
    • C07C233/01—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms
    • C07C233/45—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by carboxyl groups
    • C07C233/53—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by carboxyl groups with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by a carbon atom of a six-membered aromatic ring
    • C07C233/55—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by carboxyl groups with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by a carbon atom of a six-membered aromatic ring having the carbon atom of the carboxamide group bound to a carbon atom of an unsaturated carbon skeleton
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A61P11/06—Antiasthmatics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C275/00—Derivatives of urea, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups
    • C07C275/28—Derivatives of urea, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups having nitrogen atoms of urea groups bound to carbon atoms of six-membered aromatic rings of a carbon skeleton
    • C07C275/40—Derivatives of urea, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups having nitrogen atoms of urea groups bound to carbon atoms of six-membered aromatic rings of a carbon skeleton being further substituted by nitrogen atoms not being part of nitro or nitroso groups

Definitions

  • the present invention relates to compounds of formula (I), their preparation and use as pharmaceutical compositions as integrin antagonists, especially as ⁇ 4 ⁇ 1 and/or ⁇ 4 ⁇ 7 and/or ⁇ 9 ⁇ 1 integrin antagonists and in particular for the production of pharmaceutical compositions suitable for the inhibition or the prevention of cell adhesion and cell-adhesion mediated disorders.
  • vascular cell adhesion molecules arrest circulating leukocytes, thereby serving as the first step in their recruitment to infected or inflamed tissue sites. Subsequently, the leukocytes reaching the extravascular space interact with connective tissue cells such as fibroblasts as well as extracellular matrix proteins such as fibronectin, laminin, and collagen. Adhesion molecules on the leukocytes and on the vascular endothelium are hence essential to leukocyte migration and attractive therapeutic targets for intervention in many inflammatory disorders.
  • Leukocyte recruitment to sites of inflammation occurs in a stepwise fashion beginning with leukocyte tethering to the endothelial cells lining the blood vessels. This is followed by leukocyte rolling, activation, firm adhesion, and transmigration. A number of cell adhesion molecules involved in those four recruitment steps have been identified and characterized to date.
  • VCAM-1 vascular cell adhesion molecule 1
  • VLA-4 very late antigen 4
  • MAdCAM-1 mucosal addressin cell adhesion molecule 1
  • ⁇ 4 ⁇ 7 integrin mucosal addressin cell adhesion molecule 1
  • VCAM-1 is a member of immunoglobulin (Ig) superfamily and is one of the key regulators of leukocyte trafficking to sites of inflammation.
  • VCAM-1 along with intracellular adhesion molecule 1 (ICAM-1) and E-selectin, is expressed on inflamed endothelium activated by such cytokines as interleukin 1 (IL-1) and tumor necrosis factor ⁇ (TNF- ⁇ ), as well as by lipopolysaccharide (LPS), via nuclear factor ⁇ B (NF- ⁇ B) dependent pathway.
  • IL-1 interleukin 1
  • TNF- ⁇ B tumor necrosis factor ⁇ B
  • NF- ⁇ B nuclear factor ⁇ B
  • VCAM-1 may be involved in numerous physiological and pathological processes including myogenesis, hematopoiesis, inflammatory reactions, and the development of autoimmune disorders. Integrins VLA-4 and ⁇ 4 ⁇ 7 both function as leukocyte receptors for VCAM-1.
  • the integrin ⁇ 4 ⁇ 1 is a heterodimeric protein expressed in substantial levels on all circulating leukocytes except mature neutrophils. It regulates cell migration into tissues during inflammatory responses and normal lymphocyte trafficking.
  • VLA-4 binds to different primary sequence determinants, such as a QIDSP motif of VCAM-1 and an ILDVP sequence of the major cell type-specific adhesion site of the alternatively spliced type III connecting segment domain (CS-1) of fibronectin.
  • ⁇ 4 ⁇ 1 integrin receptor antagonists WO 96/22966, WO 97/03094, WO99/20272, WO99/26923, WO 99/33789, WO 99/37605, WO 00/00477.
  • no ⁇ -amino acids or homologues thereof with ⁇ 4 ⁇ 1 integrin receptor antagonistic activity have been described.
  • WO 98/04247 discloses cell adhesion inhibitors some of which comprise a succinic acid moiety.
  • the compounds of the present invention may also be used as ⁇ 4 ⁇ 7 or ⁇ 9 ⁇ 1 integrin antagonists.
  • An object of the present invention is to provide new succinic acid or homologues thereof derived integrin antagonists for the treatment of inflammatory, autoimmune and immune diseases.
  • the present invention therefore relates to compounds of the general formula (I) wherein
  • alkyl stands for a straight-chain or branched alkyl residue, such as methyl, ethyl, n-propyl, iso-propyl, n-pentyl. If not stated otherwise, preferred is C 1 -C 10 alkyl, very preferred is C 1 -C 6 alkyl.
  • Alkenyl and alkinyl stand for straight-chain or branched residues containing one or more double or triple bonds, e.g. vinyl, allyl, isopropinyl, ethinyl. If not stated otherwise, preferred is C 1 -C 10 alkenyl or alkinyl, very preferred is C 1 -C 6 alkenyl or alkinyl.
  • Cycloalkyl stands for a cyclic alkyl group such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl. Preferred is monocyclic C 3 -C 7 cycloalkyl.
  • Halogen in the context of the present invention stands for fluorine, chlorine, bromine or iodine. If not specified otherwise, chlorine or fluorine are preferred.
  • a 4- to 9-membered saturated, unsaturated or aromatic cyclic residue stands for a monocyclic system containing 4 to 9 ring atoms and containing 0, 1 or more double bonds, which can be attached via a carbon atom or eventually via a heteroatom within the ring, for example phenyl, thiazolyl, pyridyl, cyclopentyl.
  • Aryl stands for a monocyclic Hueckel-aromatic cyclic system containing 6 or 10 ring carbon atoms.
  • Heteroaryl stands for a monocyclic heteroaromatic system containing 4 to 9 ring atoms, which can be attached via a carbon atom or eventually via a nitrogen atom within the ring, for example, furan-2-yl, furan-3-yl, pyrrol-1-yl, pyrrol-2-yl, pyrrol-3-yl, thienyl, thiazolyl, oxazolyl, imidazolyl, triazolyl, tetrazolyl, pyridyl, pyrimidyl or pyridazinyl.
  • a saturated or unsaturated heterocyclic residue stands for a heterocyclic system containing 4 to 9 ring atoms, which can contain one or more double bonds and which can be attached via a ring carbon atom or eventually via a nitrogen atom, e.g. tetrahydrofur-2-yl, pyrrolidine-1-yl, piperidine-1-yl, piperidine-2-yl, piperidine-3-yl, piperidine-4-yl, piperazine-1-yl, piperazine-2-yl morpholine-1-yl, 1,4-diazepine-1-yl or 1,4-dihydropyridine-1-yl.
  • heteroatom stands preferably for O, S, N or P.
  • the present invention relates to compounds of general formula (I), wherein R 1 represents a phenyl ring.
  • the present invention relates to compounds of general formula (I), wherein R 1-1 represents a bond and Z represents COOR Z-1 , wherein R Z-1 has the meaning indicated above.
  • the present invention relates to compounds of general formula (I), wherein R 6 represents phenyl, which is substituted by -NHC(O)NHR 6-4 , wherein R 6-4 is substituted with methyl or trifluoromethoxy.
  • the present invention relates to compounds of general formula (I), wherein X represents a methylene group.
  • the present invention relates to compounds of general formula (I), wherein one of the substituents R 3 , R 4 , R 7 and R 8 represents phenyl, which can optionally be substituted by up to three substituents independently selected from the group C 1 -C 4 -alkyl and halogen, and the remaining substituents represent hydrogen.
  • the present invention relates to compounds of general formula (I), wherein R 1 is a 1,4-substituted phenyl ring.
  • a preferred process for preparation of compounds of general formula (I) comprises reaction of carboxylic acids of general formula (I') or activated derivatives thereof, wherein R 1 , R 2 , R 3 , R 4 , R 7 and R 8 have the above-mentioned meaning, with compounds of the general formula (I") R 6 -X-NR 5 H (I"), wherein X, R 5 and R 6 have the above-mentioned meaning, in inert solvents, which will be described in more detail in the descriptive part of the specification.
  • the compounds of the present invention show good integrin antagonistic activity. They are therefore suitable for the treatment of diseases, especially as ⁇ 4 ⁇ 1 and/or ⁇ 4 ⁇ 7 and/or ⁇ 9 ⁇ 1 integrin antagonists and in the manufacture of a medicament for the treatment or the prevention of a condition mediated by integrins and in particular for the production of pharmaceutical compositions for the inhibition or the prevention of cell adhesion and cell-adhesion mediated disorders.
  • Examples are the treatment and the prophylaxis of atherosclerosis, asthma, chronic obstructive pulmonary disease (COPD), allergies, diabetes, inflammatory bowel disease, multiple sclerosis, myocardial ischemia, rheumatoid arthritis, transplant rejection and other inflammatory, autoimmune and immune disorders.
  • COPD chronic obstructive pulmonary disease
  • integrin antagonists of the invention are useful not only for treatment of the physiological conditions discussed above, but are also useful in such activities as purification of integrins and testing for activity.
  • the compounds according to the invention can exhibit non-systemic or systemic activity, wherein the latter is preferred.
  • the active compounds can be administered, among other things, orally or parenterally, wherein oral administration is preferred.
  • parenteral administration forms of administration to the mucous membranes (i.e. buccal, lingual, sublingual, rectal, nasal, pulmonary, conjunctival or intravaginal) or into the interior of the body are particularly suitable.
  • Administration can be carried out by avoiding absorption (i.e. intracardiac, intra-arterial, intravenous, intraspinal or intralumbar administration) or by including absorption (i.e. intracutaneous, subcutaneous, percutaneous, intramuscular or intraperitoneal administration).
  • the active compounds can be administered per se or in administration forms.
  • Suitable administration forms for oral administration are, inter alia, normal and enteric-coated tablets, capsules, coated tablets, pills, granules, pellets, powders, solid and liquid aerosols, syrups, emulsions, suspensions and solutions.
  • Suitable administration forms for parenteral administration are injection and infusion solutions.
  • the active compound can be present in the administration forms in concentrations of from 0.001 - 100 % by weight; preferably the concentration of the active compound should be 0.5 - 90% by weight, i.e. quantities which are sufficient to allow the specified range of dosage.
  • the active compounds can be converted in the known manner into the above-mentioned administration forms using inert non-toxic pharmaceutically suitable auxiliaries, such as for example excipients, solvents, vehicles, emulsifiers and/or dispersants.
  • auxiliaries such as for example excipients, solvents, vehicles, emulsifiers and/or dispersants.
  • auxiliaries can be mentioned as examples: water, solid excipients such as ground natural or synthetic minerals (e.g. talcum or silicates), sugar (e.g. lactose), non-toxic organic solvents such as paraffins, vegetable oils (e.g. sesame oil), alcohols (e.g. ethanol, glycerol), glycols (e.g. polyethylene glycol), emulsifying agents, dispersants (e.g. polyvinylpyrrolidone) and lubricants (e.g. magnesium sulphate).
  • ground natural or synthetic minerals e.g. talcum or silicates
  • sugar e.g. lactose
  • non-toxic organic solvents such as paraffins, vegetable oils (e.g. sesame oil), alcohols (e.g. ethanol, glycerol), glycols (e.g. polyethylene glycol), emulsifying agents, dispersants (e.g. polyvinylpyrrolidone) and
  • tablets can of course also contain additives such as sodium citrate as well as additives such as starch, gelatin and the like.
  • Flavour enhancers or colorants can also be added to aqueous preparations for oral administration.
  • the quantity is about 0.01 to 100 mg/kg, preferably about 0.1 to 10 mg/kg of body weight.
  • compositions of the compounds of the present invention that contain an acidic moiety include addition salts formed with organic or inorganic bases.
  • the salt forming ion derived from such bases can be metal ions, e.g., aluminum, alkali metal ions, such as sodium of potassium, alkaline earth metal ions such as calcium or magnesium, or an amine salt ion, of which a number are known for this purpose.
  • Examples include ammonium salts, arylalkylamines such as dibenzylamine and N,N -dibenzylethylenediamine, lower alkylamines such as methylamine, t- butylamine, procaine, lower alkylpiperidines such as N -ethylpiperidine, cycloalkylamines such as cyclohexylamine or dicyclohexylamine, 1-adamantylamine, benzathine, or salts derived from amino acids like arginine, lysine or the like.
  • the physiologically acceptable salts such as the sodium or potassium salts and the amino acid salts can be used medicinally as described below and are preferred.
  • Pharmaceutically acceptable salts of the compounds of the present invention that contain a basic moiety include addition salts formed with organic or inorganic acids.
  • the salt forming ion derived from such acids can be halide ions or ions of natural or unnatural carboxylic or sulfonic acids, of which a number are known for this purpose. Examples include chlorides, acetates, trifluoroacetates, tartrates, or salts derived from amino acids like glycine or the like.
  • the physiologically acceptable salts such as the chloride salts, the trifluoroacetic acid salts and the amino acid salts can be used medicinally as described below and are preferred.
  • the compounds according to the invention can exist in different stereoisomeric forms, which relate to each other in an enantiomeric way (image and mirror image) or in a diastereomeric way (image different from mirror image).
  • the invention relates to the enantiomers and the diastereomers as well as their mixtures. They can be separated according to customary methods.
  • the amides (IV) By coupling of the carboxylic acid derivatives (III) with the amines (II), the amides (IV) can be obtained. Coupling of the activates carboxylic acids (IV) with the amines (V) yields the amides (VI). The removal of the protecting group PG affords carboxylic acids of type (I).
  • DCC dicyclohexyl-carbodiimid
  • EDCI 1-ethyl-3-(3'-dimethylaminopropyl)carbodiimide ⁇ HCl
  • EDCI 2-(7-aza-3-oxido-1H-1,2,3-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate.
  • the carboxyl group is preferably esterified, PG being C 1-6 -alkyl such as, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, pentyl, isopentyl, neopentyl, hexyl, a C 3-7- cycloalkyl such as, for example, cyclopropyl, cyclopropylmethyl, cyclobutyl, cyclo-pentyl, cyclohexyl, an aryl such as, for example, phenyl, benzyl, tolyl or a substituted derivative thereof.
  • PG being C 1-6 -alkyl such as, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, pentyl, isopentyl, neopentyl, he
  • Formation of the amides (IV) can take place by reacting a carboxylic acid anhydride (II) with the desired amine (III) or an acceptable salt thereof.
  • amides of type (IV) can be prepared as follows:
  • a solution of carboxylic acid anhydride in an inert solvent is stirred at r.t..
  • a non-nucleophilic base such as ethyldiisopropylamine or potassium carbonate stirring is continued at r.t. or elevated temperature.
  • the residue was redissolved in ethyl acetate, washed with aqueous acid and base, dried and evaporated. If necessary the product was purified by trituration or by flash-chromatography or used without further purification.
  • Formation of the amides (VI) can take place by reacting the respective carboxylic acids (IV) - activated by a coupling agent such as DCC and HOBt; EDCI and HOBt or HATU - with the desired amines (V) or an acceptable salt thereof.
  • a coupling agent such as DCC and HOBt; EDCI and HOBt or HATU -
  • Activated de-rivatives of the acids (IV) such as anhydrides, halides, and esters e.g. succinyl or pentafluorophenyl esters may also be employed.
  • amides (VI) can be prepared as follows:
  • Compounds of general formula (V) are commercially available, known or can be prepared by customary methods starting from known carboxylic acid derivatives.
  • Bisarylureas can be prepared by coupling of an amino phenyl acetic acid derivative and a phenylisocyanate.
  • the removal of the protecting group PG can be performed either by an acid such as trifluoroacetic acid or an base such as potassium hydroxide or lithium hydroxide, depending on the nature of PG. Reactions are carried out in aqueous, inert organic solvents such as alcohols e.g. methanol or ethanol, ethers e.g. tetrahydrofurane or dioxane or polar aprotic solvents e.g. dimethylformamide. If necessary, mixtures of the above solvents may be used.
  • the mass determinations were carried out using the electron spray ionization (ESI) method employing loop injection or split injection via a HPLC system.
  • ESI electron spray ionization
  • VCAM-1 extracellular domains 1-3
  • cDNA Complementary DNA encoding 7-domain form of VCAM-1 (GenBank accession #M60335) was obtained using Rapid-ScreenTM cDNA library panels (OriGene Technologies, Inc) at Takara Gene Analysis Center (Shiga, Japan).
  • the primers used were 5'-CCA AGG CAG AGT ACG CAA AC-3' (sense) and 5'-TGG CAG GTA TTA TTA AGG AG-3' (antisense).
  • PCR amplification of the 3-domain VCAM-1 cDNA was perform using Pfu DNA polymerase (Stratagene) with the following sets of primers: (U-VCAMd1-3) 5'-CCA TAT GGT ACC TGA TCA ATT TAA AAT CGA GAC CAC CCC AGA A-3'; (L-VCAMd1-3) 5'-CCA TAT AGC AAT CCT AGG TCC AGG GGA GAT CTC AAC AGT AAA-3'.
  • PCR cycle was 94°C for 45 sec, 55°C for 45 sec, 72°C for 2 min, repeating 15 cycles. After the purification of the PCR product, the fragment was digested with KpnI-AvrII.
  • the digested fragment was ligated into pBluescript IISK(-) (Strategene), which was linearized by digesting with KpnI-XhoI. The ligation was followed by transformation to a Dam/Dcm methylase-free E. coli strain SCS110 (Strategene) to create the donor plasmid pHH7.
  • VCAM-1 coding sequence was fused to signal peptide sequence of honeybee melittin. The resulting melittin-VCAM fusion was placed in correct orientation to the baculovirus polyhedrin promoter.
  • Baculovirus transfer vector containing first 3-domain form VCAM-1 was constructed by ligation of 0.9 kb fragment from AvrII/Klenow/BclI digests of pH7 into SalI/Klenow/BamHI digests of pMelBacB (Invitrogen).
  • Recombinant baculovirus was generated by using Bac-N-BlueTM Transfection kit (Invitrogen) according to the manufacture's instruction.
  • the recombinant virus was amplified by infection to High-FiveTM insect cells for 5 - 6 days, and virus titer was determined by plaque assay.
  • Recombinant human VCAM-1 (extracellular domains 1-3) was dissolved at 1.0 ⁇ g/ml in PBS.
  • Each well of the microtiter plates (Nalge Nunc International, Fluoronunc Cert, 437958) was coated with 100 ⁇ l of substrate or for background control with buffer alone for 15 hours at 4°C. After discarding the substrate solution, the wells were blocked using 150 ⁇ l per well of block solution (Kirkegaard Perry Laboratories, 50-61-01) for 90 minutes. The plate was washed with wash buffer containing 24 mM Tris-HCl (pH 7.4), 137 mM NaCl, 27 mM KCl and 2 mM MnCl 2 just before addition of the assay.
  • Ramos cells (American Type Culture Collection, Clone CRL-1596) were cultured in RPMI 1640 medium (Nikken Bio Medical Laboratory, CM1101) supplemented with 10% fetal bovine serum (Hyclone, A-1119-L), 100 U/ml penicilin (Gibco BRL, 15140-122) and 100 ⁇ g/ml streptomycin (Gibco BRL, 15140-122) in a humidified incubator at 37°C with 5% CO 2 .
  • Ramos cells were incubated with phosphate balanced solution (PBS, Nissui, 05913) containing 25 ⁇ M of 5(-and -6)-carboxyfluorescein diacetate, succinimidyle ester (CFSE, Dojindo Laboratories, 345-06441) for 20 min at room temperature while gently swirling every 5 min. After centrifugation at 1000 rpm for 5 min, the cell pellet was resuspended with adhesion assay buffer at a cell density of 4 x 10 6 cells/ml.
  • PBS phosphate balanced solution
  • CFSE succinimidyle ester
  • the adhesion assay buffer was composed of 24 mM Tris-HCl (pH 7.4), 137 mM NaCl, 27 mM KCl, 4 mM glucose, 0.1 % bovine serum albumin (BSA, Sigma, A9647) and 2 mM MnCl 2 .
  • the assay solution containing each test compounds or 5 ⁇ g/ml anti-CD49d monoclonal antibody (Immunotech, 0764) was transferred to the VCAM-1 coated plates.
  • the final concentration of each test compounds was 5 ⁇ M, 10 ⁇ M or various concentrations ranging from 0.0001 ⁇ M to 10 ⁇ M using a standard 5-point serial dilution.
  • the assay solution containing the labeled Ramos cells was transferred to the VCAM-1 coated plates at a cell density of 2 x 10 5 cells per well and incubated for 1 hour at 37°C. The non-adherent cells were removed by washing the plates 3 times with wash buffer.
  • the adherent cells were broken by addition of 1 % Triton X-100 (Nacalai Tesque, 355-01). Released CFSC was quantified fluorescence measurement in a fluorometer (Wallac, ARVO 1420 multilabel counter).
  • FTB the total fluorescent intensity from VCAM-1 coated wells without test compound
  • FBG the fluorescent intensity from wells with anti-CD49d monoclonal antibody
  • FTS the fluorescent intensity from wells containing the test compound of this invention.

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Abstract

The present invention relates to compounds of the general formula (I), processes for their preparation, pharmaceutical compositions containing them as well as their use for the production of pharmaceutical compositions for the treatment of inflammatory diseases

Description

  • The present invention relates to compounds of formula (I),
    Figure imgb0001
     their preparation and use as pharmaceutical compositions as integrin antagonists, especially as α4β1 and/or α4β7 and/or α9β1 integrin antagonists and in particular for the production of pharmaceutical compositions suitable for the inhibition or the prevention of cell adhesion and cell-adhesion mediated disorders.
  • Adhesive interactions between the leukocytes and endothelial cells play a critical role in leukocyte trafficking to sites of inflammation. These events are essential for normal host defense against pathogens and repair of tissue damage, but can also contribute to the pathology of a variety of inflammatory and autoimmune disorders. Indeed, eosinophil and T cell infiltration into the tissue is known as a cardinal feature of allergic inflammation such as asthma.
  • The interaction of circulating leukocytes with adhesion molecules on the luminal surface of blood vessels appears to modulate leukocyte transmigration. These vascular cell adhesion molecules arrest circulating leukocytes, thereby serving as the first step in their recruitment to infected or inflamed tissue sites. Subsequently, the leukocytes reaching the extravascular space interact with connective tissue cells such as fibroblasts as well as extracellular matrix proteins such as fibronectin, laminin, and collagen. Adhesion molecules on the leukocytes and on the vascular endothelium are hence essential to leukocyte migration and attractive therapeutic targets for intervention in many inflammatory disorders.
  • Leukocyte recruitment to sites of inflammation occurs in a stepwise fashion beginning with leukocyte tethering to the endothelial cells lining the blood vessels. This is followed by leukocyte rolling, activation, firm adhesion, and transmigration. A number of cell adhesion molecules involved in those four recruitment steps have been identified and characterized to date. Among them, the interaction between vascular cell adhesion molecule 1 (VCAM-1) and very late antigen 4 (VLA-4, α4β1 integrin), as well as the interaction between mucosal addressin cell adhesion molecule 1 (MAdCAM-1) and α4β7 integrin, has been shown to mediate the tethering, rolling, and adhesion of lymphocytes and eosinophils, but not neutrophils, to endothelial cells under a physiologic flow condition. This suggests that the VCAM-1 / VLA-4 and/or MAdCAM-1 / α4β7 integrin mediated interactions could predominantly mediate a selective recruitment of leukocyte subpopulations in vivo. The inhibition of this interaction is a point of departure for therapeutic intervention (A. J. Wardlaw, J. Allergy Clin. Immunol. 1999, 104, 917-26).
  • VCAM-1 is a member of immunoglobulin (Ig) superfamily and is one of the key regulators of leukocyte trafficking to sites of inflammation. VCAM-1, along with intracellular adhesion molecule 1 (ICAM-1) and E-selectin, is expressed on inflamed endothelium activated by such cytokines as interleukin 1 (IL-1) and tumor necrosis factor α (TNF-α), as well as by lipopolysaccharide (LPS), via nuclear factor κB (NF-κB) dependent pathway. However, these molecules are not expressed on resting endothelium. Cell adhesion mediated by VCAM-1 may be involved in numerous physiological and pathological processes including myogenesis, hematopoiesis, inflammatory reactions, and the development of autoimmune disorders. Integrins VLA-4 and α4β7 both function as leukocyte receptors for VCAM-1.
  • The integrin α4β1 is a heterodimeric protein expressed in substantial levels on all circulating leukocytes except mature neutrophils. It regulates cell migration into tissues during inflammatory responses and normal lymphocyte trafficking. VLA-4 binds to different primary sequence determinants, such as a QIDSP motif of VCAM-1 and an ILDVP sequence of the major cell type-specific adhesion site of the alternatively spliced type III connecting segment domain (CS-1) of fibronectin.
  • In vivo studies with neutralizing monoclonal antibodies and inhibitor peptides have demonstrated a critical role for α4 integrins interaction in leukocyte-mediated inflammation. Blocking of VLA-4/ligand interactions, thus, holds promise for therapeutic intervention in a variety of inflammatory, autoimmune and immune diseases (Zimmerman, C.; Exp. Opin. Ther. Patents 1999, 9, 129-133).
  • Furthermore, compounds containing a bisarylurea moiety as a substituent were disclosed as α4β1 integrin receptor antagonists: WO 96/22966, WO 97/03094, WO99/20272, WO99/26923, WO 99/33789, WO 99/37605, WO 00/00477. However, no β-amino acids or homologues thereof with α4β1 integrin receptor antagonistic activity have been described.
  • WO 98/04247 discloses cell adhesion inhibitors some of which comprise a succinic acid moiety.
  • Further to their α4β1 integrin antagonistic activity, the compounds of the present invention may also be used as α4β7 or α9β1 integrin antagonists.
  • An object of the present invention is to provide new succinic acid or homologues thereof derived integrin antagonists for the treatment of inflammatory, autoimmune and immune diseases.
  • The present invention therefore relates to compounds of the general formula (I)
    Figure imgb0002
     wherein
    • R1
    represents a 4- to 9-membered saturated, unsaturated or aromatic cyclic residue,
    which can contain 0 to 3 heteroatoms selected independently from the group N, S and O,
    and wherein R1 is substituted by -R1-1-Z,
    wherein
    R1-1
    represents a bond, -O-, -S-, NR1-2, C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C6 or C10 aryl, C3-C7 cycloalkyl or a 4-9-membered saturated or unsaturated heterocyclic residue containing up to 3 heteroatoms selected from the group oxygen, nitrogen or sulfur,
    wherein R1-1 can optionally be substituted by 1 to 2 substituents selected from the group R1-3,
    wherein R1-2 can optionally be hydrogen, C1-C10 alkyl, C2-C10 alkenyl or C2-C10 alkynyl, and
    wherein R1-3 represents hydrogen, C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C6 or C10 aryl, C3-C7 cycloalkyl or a 4-9-membered saturated or unsaturated heterocyclic residue containing up to 3 heteroatoms selected from the group oxygen, nitrogen or sulfur,
    Z
    represents -C(O)ORZ-1, -C(O)NRZ-2RZ-3, -SO2NRZ-2RZ-3, -SO(ORZ-1), -SO2(ORZ-1), -P(O)RZ-1(ORZ-3) or -PO(ORZ-1)(ORZ-3),
    wherein RZ-2 is hydrogen, C1-C4 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C6 or C10 aryl, -C(O)RZ-4 or -SO2RZ-4,
    wherein RZ-4 is C1-C4 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C6 or C10 aryl,
    RZ-1 and RZ-3
    are independently selected from the group hydrogen, C1-C4 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C6 or C10 aryl or benzyl,
    wherein RZ-1 and RZ-3 can optionally be substituted by 1 to 3 substituents selected from the group C1-C4 alkyl, C1-C4 alkyloxy, halogen, nitro, cyano,

    and wherein R1 can optionally be substituted by 0 to 2 substituents R1-4, halogen, nitro, amino, cyano and oxo,
    wherein
    R1-4
    is selected from the group C1-C4 alkyl, C1-C4 alkyloxy, phenyl, phenoxy, phenylamino, C3-C6 cycloalkyl,
    R2
    represents hydrogen, C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C6 or C10 aryl or C3-C7 cycloalkyl,

    wherein R2 can optionally be substituted by 1 to 3 radicals independently selected from the group C1-C4 alkyl, trifluormethyl, trifluormethoxy, halogen, cyano, nitro or oxo,
    R3
    represents hydrogen, C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C6 or C10 aryl, C3-C7 cycloalkyl or a 4-9-membered saturated or unsaturated heterocyclic residue containing up to 2 heteroatoms selected from the group oxygen, nitrogen or sulfur,

    wherein R3 can optionally be substituted by 1 to 3 radicals R3-1,
    wherein R3-1 represents C1-C4 alkyl, trifluormethyl, trifluormethoxy, -OR3-2, -SR3-2, NR3-3R3-4, -C(O)R3-2, S(O)R3-2, -SO2R3-2, -OC(O)R3-2, -C(O)NR3-3R3-4, -NR3-2C(O)R3-3, -SO2NR3-3R3-4, NR3-2SO2R3-3, -NR3-2C(O)NR3-3R3-4, -NR3-2C(O)OR3-3, -OC(O)NR3-3R3-4, -CO2R3-5, halogen, cyano, nitro or oxo,
    wherein R3-2 represents hydrogen, C1-C4 alkyl, C3-C6 cycloalkyl, C6 or C10 aryl,
    wherein R3-3 and R3-4 are independently selected from the group hydrogen, C1-C4 alkyl, C3-C6 cycloalkyl, C6 or C10 aryl or benzyl,
    and wherein R3-5 represents C1-C4 alkyl, C3-C6 cycloalkyl, C6 or C10 aryl,
    R4
    represents hydrogen, C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C6 or C10 aryl, C3-C7 cycloalkyl or a 4-9-membered saturated or unsaturated heterocyclic residue containing up to 2 heteroatoms selected from the group oxygen, nitrogen or sulfur,

    wherein R4 can optionally be substituted by 1 to 3 radicals R4-1,
    wherein R4-1 represents C1-C4 alkyl, trifluormethyl, trifluormethoxy, -OR4-2, -SR4-2, NR4-3R4-4, -C(O)R4-2, S(O)R4-2, -SO2R4-2, -OC(O)R4-2, -C(O)NR4-3R4-4, -NR4-2C(O)R4-3, -SO2NR4-3R4-4, NR4-2SO2R4-3, -NR4-2C(O)NR4-3R4-4, -NR4-2C(O)OR4-3, -OC(O)NR4-3R4-4, -CO2R4-5, halogen, cyano, nitro or oxo,
    wherein R4-2 represents hydrogen, C1-C4 alkyl, C3-C6 cycloalkyl, C6 or C10 aryl,
    wherein R4-3 and R4-4 are independently selected from the group hydrogen, C1-C4 alkyl, C3-C6 cycloalkyl, C6 or C10 aryl or benzyl,
    and wherein R4-5 represents C1-C4 alkyl, C3-C6 cycloalkyl, C6 or C10 aryl
    or
    R3 and R4
    together with the carbon atom to which they are attached form a 4-7-membered saturated or unsaturated ring containing up to 2 heteroatoms selected from the group oxygen, nitrogen or sulfur, which can optionally be substituted by 1 to 2 substituents selected from the group C1-C4 alkyl, phenyl, benzyl, C3-C7 cycloalkyl, C1-C4 alkyloxy, halogen, nitro, cyano, oxo and which can be fused with a 3-7 membered homocyclic or heterocyclic, saturated, unsaturated or aromatic ring,
    R5
    represents hydrogen, C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C6 or C10 aryl or C3-C7 cycloalkyl,

    wherein R5 can optionally up to threefoldedly be substituted by C1-C4 alkyl, trifluormethyl, trifluormethoxy, halogen, cyano, nitro or oxo,
    R6
    represents phenyl or a 5- to 6-membered aromatic heterocyclic residue containing up to 3 heteroatoms independently selected from the group oxygen, nitrogen or sulfur,

    wherein R6 is substituted by -NR6-2C(O)NR6-3R6-4 or -NR6-2C(S)NR6-3R6-4 and can furthermore optionally be substituted by halogen,
    wherein R6-2 and R6-3 are independently selected from the group hydrogen or C1-C4 alkyl, or together form a group
    Figure imgb0003
    and wherein R6-4 represents phenyl,
    wherein R6-4 can optionally be substituted by 1-2 substituents selected from the group C1-C4 alkyl, C1-C4 alkyloxy, halogen, nitro, trifluoromethyl, trifluoromethoxy or cyano,
    or
    R6
    represents a group
    Figure imgb0004
    wherein R6-1 represents a substituent selected from the group hydrogen, C1-C4 alkyl, C1-C4 alkyloxy, halogen, nitro, trifluoromethyl, trifluoromethoxy or cyano,
    and
    wherein R6-5 represents a substituent selected from the group hydrogen, C1-C4 alkyl, C1-C4 alkyloxy, halogen, nitro, trifluoromethyl, trifluoromethoxy or cyano,
    R7
    represents hydrogen, C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C6 or C10 aryl, C3-C7 cycloalkyl or a 4-9-membered saturated or unsaturated heterocyclic residue containing up to 2 heteroatoms selected from the group oxygen, nitrogen or sulfur,

    wherein R7 can optionally be substituted by 1 to 3 radicals R7-1,
    wherein R7-1 represents C1-C4 alkyl, trifluormethyl, trifluormethoxy, -OR7-2, -SR7-2, NR7-3R7-4, -C(O)R7-2, S(O)R7-2, -SO2R7-2, -OC(O)R7-2, -C(O)NR7-3R7-4, -NR7-2C(O)R7-3, -SO2NR7-3R7-4, NR7-2SO2R7-3, -NR7-2C(O)NR7-3R7-4, -NR7-2C(O)OR7-3, -OC(O)NR7-3R7-4, -CO2R7-5, halogen, cyano, nitro or oxo,
    wherein R7-2 represents hydrogen, C1-C4 alkyl, C3-C6 cycloalkyl, C6 or C10 aryl,
    wherein R7-3 and R7-4 are independently selected from the group hydrogen, C1-C4 alkyl, C3-C6 cycloalkyl, C6 or C10 aryl or benzyl,
    and wherein R7-5 represents C1-C4 alkyl, C3-C6 cycloalkyl, C6 or C10 aryl
    R8
    represents hydrogen, C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C6 or C10 aryl, C3-C7 cycloalkyl or a 4-9-membered saturated or unsaturated heterocyclic residue containing up to 2 heteroatoms selected from the group oxygen, nitrogen or sulfur,

    wherein R8 can optionally be substituted by 1 to 3 radicals R8-1,
    wherein R8-1 represents C1-C4 alkyl, trifluormethyl, trifluormethoxy, -OR8-2, -SR8-2, NR8-3R8-4, -C(O)R8-2, S(O)R8-2, -SO2R8-2, -OC(O)R8-2, -C(O)NR8-3R8-4, -NR8-2C(O)R8-3, -SO2NR8-3R8-4, NR8-2SO2R8-3, -NR8-2C(O)NR8-3R8-4, -NR8-2C(O)OR8-3, -OC(O)NR8-3R8-4, -CO2R8-5, halogen, cyano, nitro or oxo,
    wherein R8-2 represents hydrogen, C1-C4 alkyl, C3-C6 cycloalkyl, C6 or C10 aryl,
    wherein R8-3 and R8-4 are independently selected from the group hydrogen, C1-C4 alkyl, C3-C6 cycloalkyl, C6 or C10 aryl or benzyl,
    and wherein R8-5 represents C1-C4 alkyl, C3-C6 cycloalkyl, C6 or C10 aryl
    or
    R7 and R8
    together form a 4-7-membered saturated or unsaturated ring containing up to 2 heteroatoms selected from the group oxygen, nitrogen or sulfur, which can optionally be substituted by 1 to 2 substituents selected from the group C1-C4 alkyl, phenyl, benzyl, C3-C7 cycloalkyl, C1-C4 alkyloxy, halogen, nitro, cyano, oxo and which can be fused with a 3-7 membered homocyclic or heterocyclic, saturated, unsaturated or aromatic ring,
    X
    represents bond or (-CRX-1RX-2-)n,

    wherein RX-1 and RX-2 can be independently selected from the group hydrogen, C1-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl,
    wherein RX-1 and RX-2 can optionally independently be substituted by 1 to 2 substituents selected from the group C1-C4 alkyl, phenyl, benzyl, C3-C7 cycloalkyl, C1-C4 alkyloxy, halogen, nitro, cyano, oxo,
    and wherein n is an integer 0 or 1,
    and pharmaceutically acceptable salts thereof.
  • In the context of the present invention alkyl stands for a straight-chain or branched alkyl residue, such as methyl, ethyl, n-propyl, iso-propyl, n-pentyl. If not stated otherwise, preferred is C1-C10 alkyl, very preferred is C1-C6 alkyl.
  • Alkenyl and alkinyl stand for straight-chain or branched residues containing one or more double or triple bonds, e.g. vinyl, allyl, isopropinyl, ethinyl. If not stated otherwise, preferred is C1-C10 alkenyl or alkinyl, very preferred is C1-C6 alkenyl or alkinyl.
  • Cycloalkyl stands for a cyclic alkyl group such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl. Preferred is monocyclic C3-C7 cycloalkyl.
  • Halogen in the context of the present invention stands for fluorine, chlorine, bromine or iodine. If not specified otherwise, chlorine or fluorine are preferred.
  • A 4- to 9-membered saturated, unsaturated or aromatic cyclic residue stands for a monocyclic system containing 4 to 9 ring atoms and containing 0, 1 or more double bonds, which can be attached via a carbon atom or eventually via a heteroatom within the ring, for example phenyl, thiazolyl, pyridyl, cyclopentyl.
  • Aryl stands for a monocyclic Hueckel-aromatic cyclic system containing 6 or 10 ring carbon atoms.
  • Heteroaryl stands for a monocyclic heteroaromatic system containing 4 to 9 ring atoms, which can be attached via a carbon atom or eventually via a nitrogen atom within the ring, for example, furan-2-yl, furan-3-yl, pyrrol-1-yl, pyrrol-2-yl, pyrrol-3-yl, thienyl, thiazolyl, oxazolyl, imidazolyl, triazolyl, tetrazolyl, pyridyl, pyrimidyl or pyridazinyl.
  • A saturated or unsaturated heterocyclic residue stands for a heterocyclic system containing 4 to 9 ring atoms, which can contain one or more double bonds and which can be attached via a ring carbon atom or eventually via a nitrogen atom, e.g. tetrahydrofur-2-yl, pyrrolidine-1-yl, piperidine-1-yl, piperidine-2-yl, piperidine-3-yl, piperidine-4-yl, piperazine-1-yl, piperazine-2-yl morpholine-1-yl, 1,4-diazepine-1-yl or 1,4-dihydropyridine-1-yl.
  • If not specified otherwise, in the context of the present invention heteroatom stands preferably for O, S, N or P.
  • Compounds according to the invention, wherein one of the substituents R3, R4, R7 and R8 represents phenyl, which can optionally be substituted by up to three substituents independently selected from the group C1-C4-alkyl and halogen, and the remaining substituents represent hydrogen are such that have one single phenyl group bound to any one of the chain carbon atoms to which these substituents are attached.
  • In a preferred embodiment, the present invention relates to compounds of general formula (I), wherein R1 represents a phenyl ring.
  • In another preferred embodiment, the present invention relates to compounds of general formula (I), wherein R1-1 represents a bond and Z represents COORZ-1, wherein RZ-1 has the meaning indicated above.
  • In yet another preferred embodiment, the present invention relates to compounds of general formula (I), wherein R6 represents phenyl, which is substituted by -NHC(O)NHR6-4, wherein R6-4 is substituted with methyl or trifluoromethoxy.
  • In yet another preferred embodiment, the present invention relates to compounds of general formula (I), wherein X represents a methylene group.
  • In yet another preferred embodiment, the present invention relates to compounds of general formula (I), wherein one of the substituents R3, R4, R7 and R8 represents phenyl, which can optionally be substituted by up to three substituents independently selected from the group C1-C4-alkyl and halogen, and the remaining substituents represent hydrogen.
  • In yet another preferred embodiment, the present invention relates to compounds of general formula (I), wherein R1 is a 1,4-substituted phenyl ring.
  • A preferred process for preparation of compounds of general formula (I) has also been found, which comprises reaction of carboxylic acids of general formula (I')
    Figure imgb0005
    or activated derivatives thereof, wherein
    R1, R2, R3, R4, R7 and R8 have the above-mentioned meaning,
    with compounds of the general formula (I")

            R6-X-NR5H     (I"),

    wherein
    X, R5 and R6 have the above-mentioned meaning,
    in inert solvents, which will be described in more detail in the descriptive part of the specification.
  • Surprisingly, the compounds of the present invention show good integrin antagonistic activity. They are therefore suitable for the treatment of diseases, especially as α4β1 and/or α4β7 and/or α9β1 integrin antagonists and in the manufacture of a medicament for the treatment or the prevention of a condition mediated by integrins and in particular for the production of pharmaceutical compositions for the inhibition or the prevention of cell adhesion and cell-adhesion mediated disorders. Examples are the treatment and the prophylaxis of atherosclerosis, asthma, chronic obstructive pulmonary disease (COPD), allergies, diabetes, inflammatory bowel disease, multiple sclerosis, myocardial ischemia, rheumatoid arthritis, transplant rejection and other inflammatory, autoimmune and immune disorders.
  • The integrin antagonists of the invention are useful not only for treatment of the physiological conditions discussed above, but are also useful in such activities as purification of integrins and testing for activity.
  • For the treatment of the above-mentioned diseases, the compounds according to the invention can exhibit non-systemic or systemic activity, wherein the latter is preferred. To obtain systemic activity the active compounds can be administered, among other things, orally or parenterally, wherein oral administration is preferred.
  • For parenteral administration, forms of administration to the mucous membranes (i.e. buccal, lingual, sublingual, rectal, nasal, pulmonary, conjunctival or intravaginal) or into the interior of the body are particularly suitable. Administration can be carried out by avoiding absorption (i.e. intracardiac, intra-arterial, intravenous, intraspinal or intralumbar administration) or by including absorption (i.e. intracutaneous, subcutaneous, percutaneous, intramuscular or intraperitoneal administration).
  • For the above purpose the active compounds can be administered per se or in administration forms.
  • Suitable administration forms for oral administration are, inter alia, normal and enteric-coated tablets, capsules, coated tablets, pills, granules, pellets, powders, solid and liquid aerosols, syrups, emulsions, suspensions and solutions. Suitable administration forms for parenteral administration are injection and infusion solutions.
  • The active compound can be present in the administration forms in concentrations of from 0.001 - 100 % by weight; preferably the concentration of the active compound should be 0.5 - 90% by weight, i.e. quantities which are sufficient to allow the specified range of dosage.
  • The active compounds can be converted in the known manner into the above-mentioned administration forms using inert non-toxic pharmaceutically suitable auxiliaries, such as for example excipients, solvents, vehicles, emulsifiers and/or dispersants.
  • The following auxiliaries can be mentioned as examples: water, solid excipients such as ground natural or synthetic minerals (e.g. talcum or silicates), sugar (e.g. lactose), non-toxic organic solvents such as paraffins, vegetable oils (e.g. sesame oil), alcohols (e.g. ethanol, glycerol), glycols (e.g. polyethylene glycol), emulsifying agents, dispersants (e.g. polyvinylpyrrolidone) and lubricants (e.g. magnesium sulphate).
  • In the case of oral administration tablets can of course also contain additives such as sodium citrate as well as additives such as starch, gelatin and the like. Flavour enhancers or colorants can also be added to aqueous preparations for oral administration.
  • For the obtainment of effective results in the case of parenteral administration it has generally proven advantageous to administer quantities of about 0.001 to 100 mg/kg, preferably about 0.01 to 1 mg/kg of body weight. In the case of oral administration the quantity is about 0.01 to 100 mg/kg, preferably about 0.1 to 10 mg/kg of body weight.
  • It may nevertheless be necessary to use quantities other than those mentioned above, depending on the body weight concerned, the method of administration, the individual response to the active compound, the type of preparation and the time or interval of administration.
  • Pharmaceutically acceptable salts of the compounds of the present invention that contain an acidic moiety include addition salts formed with organic or inorganic bases. The salt forming ion derived from such bases can be metal ions, e.g., aluminum, alkali metal ions, such as sodium of potassium, alkaline earth metal ions such as calcium or magnesium, or an amine salt ion, of which a number are known for this purpose. Examples include ammonium salts, arylalkylamines such as dibenzylamine and N,N-dibenzylethylenediamine, lower alkylamines such as methylamine, t-butylamine, procaine, lower alkylpiperidines such as N-ethylpiperidine, cycloalkylamines such as cyclohexylamine or dicyclohexylamine, 1-adamantylamine, benzathine, or salts derived from amino acids like arginine, lysine or the like. The physiologically acceptable salts such as the sodium or potassium salts and the amino acid salts can be used medicinally as described below and are preferred.
  • Pharmaceutically acceptable salts of the compounds of the present invention that contain a basic moiety include addition salts formed with organic or inorganic acids. The salt forming ion derived from such acids can be halide ions or ions of natural or unnatural carboxylic or sulfonic acids, of which a number are known for this purpose. Examples include chlorides, acetates, trifluoroacetates, tartrates, or salts derived from amino acids like glycine or the like. The physiologically acceptable salts such as the chloride salts, the trifluoroacetic acid salts and the amino acid salts can be used medicinally as described below and are preferred.
  • These and other salts which are not necessarily physiologically acceptable are useful in isolating or purifying a product acceptable for the purposes described below.
  • The compounds according to the invention can exist in different stereoisomeric forms, which relate to each other in an enantiomeric way (image and mirror image) or in a diastereomeric way (image different from mirror image). The invention relates to the enantiomers and the diastereomers as well as their mixtures. They can be separated according to customary methods.
    • General compound synthesis
  • The synthesis of compounds according to the general formula (I) can be illustrated by the following scheme 1:
    Figure imgb0006
  • By coupling of the carboxylic acid derivatives (III) with the amines (II), the amides (IV) can be obtained. Coupling of the activates carboxylic acids (IV) with the amines (V) yields the amides (VI). The removal of the protecting group PG affords carboxylic acids of type (I).
  • In the above scheme the depicted ring in formulas (I), (II), (IV), and (VI) represents a cyclic moiety formed by R1. Activated carboxylic acids derivatives of this type are known to the person skilled in the art and are described in detail in standard textbooks such as, for example in (i) Houben-Weyl, Methoden der organischen Chemie [Methods of Organic Chemistry], Georg Thieme Verlag, Stuttgart or (ii) Comprehensive Organic Synthesis, Ed. B. M. Trost, Pergamon Press, Oxford, 1991. The carboxylic acid is preferably activated as, such as, for example, AG = 1-hydroxy-1H-benzotriazol and a coupling agents such as, for example, dicyclohexyl-carbodiimid (DCC), 1-ethyl-3-(3'-dimethylaminopropyl)carbodiimide×HCl (EDCI), 2-(7-aza-3-oxido-1H-1,2,3-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate. Other activated carboxylic acid derivatives such as, for example symmetric anhydrides, mixed anhydrides, N-carboxy anhydrides, halides, or further activated esters e.g. succinyl or pentafluorophenyl esters may also be employed.
  • In the above scheme PG stand for a suitable protecting group of the carboxyl group or COOPG stand for the carboxylic group attached to a polymeric resin suitable for solid phase synthesis. Protecting groups of this type are known to the person skilled in the art and are described in detail in T. W. Greene, P. G. Wuts, Protective Groups in Organic Synthesis, 3rd ed., John Wiley, New York, 1999. The carboxyl group is preferably esterified, PG being C1-6-alkyl such as, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, pentyl, isopentyl, neopentyl, hexyl, a C3-7-cycloalkyl such as, for example, cyclopropyl, cyclopropylmethyl, cyclobutyl, cyclo-pentyl, cyclohexyl, an aryl such as, for example, phenyl, benzyl, tolyl or a substituted derivative thereof.
    • Step A
  • Formation of the amides (IV) can take place by reacting a carboxylic acid anhydride (II) with the desired amine (III) or an acceptable salt thereof.
  • For example, amides of type (IV) can be prepared as follows:
    • Anhydride procedure
  • A solution of carboxylic acid anhydride in an inert solvent is stirred at r.t.. After addition of the amine and a non-nucleophilic base such as ethyldiisopropylamine or potassium carbonate stirring is continued at r.t. or elevated temperature. After evaporation, the residue was redissolved in ethyl acetate, washed with aqueous acid and base, dried and evaporated. If necessary the product was purified by trituration or by flash-chromatography or used without further purification.
  • Compounds of general formula (II) are commercially available, known or can be prepared by customary methods starting from commercially available precursors.
  • Compounds of general formula (III) are commercially available, known or can be prepared by customary methods starting from known carboxylic acid derivatives.
    • Step B
  • Formation of the amides (VI) can take place by reacting the respective carboxylic acids (IV) - activated by a coupling agent such as DCC and HOBt; EDCI and HOBt or HATU - with the desired amines (V) or an acceptable salt thereof. Activated de-rivatives of the acids (IV) such as anhydrides, halides, and esters e.g. succinyl or pentafluorophenyl esters may also be employed.
  • For example, amides (VI) can be prepared as follows:
  • A solution of carboxylic acid, 1-hydroxy-1H-benzotriazol (HOBt) and 1-ethyl-3-(3'-dimethylaminopropyl)carbodiimide×HCl (EDCI) in an inert solvent is stirred at r.t.. After addition of the amine and a non-nucleophilic base such as ethyldiisopropylamine or potassium carbonate stirring is continued at r.t. or elevated temperature. After evaporation, the residue was redissolved in ethyl acetate, washed with aqueous acid and base, dried and evaporated. If necessary the product was purified by trituration or by flash-chromatography or used without further purification.
  • Compounds of general formula (V) are commercially available, known or can be prepared by customary methods starting from known carboxylic acid derivatives. Bisarylureas can be prepared by coupling of an amino phenyl acetic acid derivative and a phenylisocyanate.
    • Step C
  • The removal of the protecting group PG can be performed either by an acid such as trifluoroacetic acid or an base such as potassium hydroxide or lithium hydroxide, depending on the nature of PG. Reactions are carried out in aqueous, inert organic solvents such as alcohols e.g. methanol or ethanol, ethers e.g. tetrahydrofurane or dioxane or polar aprotic solvents e.g. dimethylformamide. If necessary, mixtures of the above solvents may be used.
    • Examples Abbreviations
    • AcOH
    acetic acid
    Boc
    tert-butyloxycarbonyl
    DCC
    dicyclohexylcarbodiimid
    DCM
    dichloromethane
    DIPEA
    diisopropylethylamine
    EDCI
    1-ethyl-3-(3'-dimethylaminopropyl)carbodiimide×HCl
    eq.
    equivalents
    EtOAc
    ethyl acetate
    FC
    flash chromatography
    GC
    gas chromatography
    HATU
    2-(7-aza-3-oxido-1H-1,2,3-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate
    HOBt
    N-hydroxybenzotriazole monohydrate
    HPLC
    high performance liquid chromatography
    ICAM-1
    intracellular adhesion molecule 1
    IL-1
    interleukin 1
    LPS
    lipopolysaccharide
    MAdCAM-1
    mucosal addressin cell adhesion molecule 1
    MeOH
    methanol
    MeCN
    acetonitril
    min.
    minutes
    M.p.
    melting point
    NF-κB
    nuclear factor κB
    NMR
    nuclear magnetic resonance
    n.d.
    not determined
    PE
    light petroleum (b.p. 40-60 °C)
    r.t.
    room temperature
    Rf
    TLC: Rf value = distance spot traveled / distance solvent front traveled
    TFA
    trifluoroacetic acid
    THF
    tetrahydrofurane
    TLC
    thin layer chromatography
    TNF-α
    tumor necrosis factor α
    tR
    retention time determined by HPLC
    VCAM-1
    vascular cell adhesion molecule 1
    VLA-4
    very late antigen 4 (α4β1 integrin)
    General remarks
  • In the examples below, all quantitative data, if not stated otherwise, relate to percentages by weight.
  • Flash chromatography was carried out on silica gel 60, 40-63µm (E. Merck, Darmstadt, Germany).
  • Thin layer chromatography was carried out, employing silica gel 60 F254 coated aluminum sheets (E. Merck, Darmstadt, Germany) with the mobile phase indicated.
  • Melting points were determined in open capillaries and are not corrected.
  • The mass determinations were carried out using the electron spray ionization (ESI) method employing loop injection or split injection via a HPLC system.
    • Precursor synthesis Example I: N-(4-Aminophenyl)-N'-(2-methylphenyl)urea
  • Figure imgb0007
  • 2-Methylphenylisocyanate (24.6 g, 184.9 mmol) was added dropwise at 0°C to a solution of 1,4-diamino benzene (20.00 g, 184.9 mmol) in 1000 mL EtOAc. After stirring for 2 h at r.t. the product was collected by filtration (42.7 g, 177.0 mmol). M.p. >300°C;
    TLC (PE/EtOAc 1/4) Rf 0.32; 1H-NMR (400 MHz, D6-DMSO) δ 2.10 (s, 3H); 4.76 (s, 2H); 6.59 (mc, 2H); 6.89 (mc, 1H); 7.07-7.15 (m, 4H); 7.73 (s, 1H); 7.85 (mc, 2H); 8.50 (s, 1H).
    • Example II: tert-Butyl 4-({[(2-methylphenyl)amino]carbonyl}amino)benzylcarbamate
  • Figure imgb0008
  • 2-Methylphenylisocyanate (7.57 g, 59.83 mmol) was added dropwise at 0°C to a solution of (4-amino-benzyl)-carbamic acid tert-butyl ester (13.30 g, 59.83 mmol, prepared analoguous to: Moloney, Gerard P.; Martin, Graeme R.; Mathews, Neil; Milne, Aynsley; Hobbs, Heather; et al. J. Med. Chem. 1999, 42, 2504 - 2526) in 120 mL DCM. The reaction was heated under reflux for 16 h, cooled to r.t. and the precipitated product was collected by filtration and dried in vacuum (19.20 g, 54.00 mmol). M.p. 200-202 °C; TLC (PE/EtOAc 1/1) Rf 0.65; 1H NMR (400 MHz, D6-DMSO) δ 1.39 (s, 9H); 2.24 (s, 3H); 4.06 (d, J=6 Hz, 2H); 6.93 (mc, 1H); 7.12-7.17 (m, 4); 7.32 (mc, 1H); 7.40 (mc, 2H); 7.85 (mc, 1H); 7.90 (s, 1H); 8.98 (s, 1H).
    • Example III: N-[4-(Aminomethyl)phenyl]-N'-(2-methylphenyl)urea
  • Figure imgb0009
  • To a solution of tert-butyl 4-({[(2-methylphenyl)amino]carbonyl}amino)benzylcarbamate (2.00 g, 5.63 mmol) in CH2Cl2 (120 mL) TFA (36 mL) was added at 0°C and stirred for 2 h at r.t.. The reaction mixture was evaporated and the product was collected (2.72 g, TFA salt). M.p. 142-143°C; TLC (PE/EtOAc 3/2) Rf 0.14; 1H NMR (400 MHz, D6-DMSO) δ 2.24 (s, 3H); 3.97 (q, J=5 Hz, 2H); 6.96 (mc, 1H); 7.13-7.19 (m, 2); 7.36 (mc, 2H); 7.51 (mc, 2H); 7.81 (mc, 2H); 8.06 (s, 1H); 8.08 (s, 3H); 9.23 (s, 1H).
    • Compound synthesis, Example 1 Step A Example IV: 4-{[4-(ethoxycarbonyl)phenyl]amino}-4-oxo-2-phenylbutanoic acid
  • Figure imgb0010
  • Ethyl 4-aminobenzoate (910 mg, 5.50 mmol) and phenyl succinic anhydride (1070 mg, 6.00 mmol) were dissolved in 40mL abs. CH3CN and stirred overnight at r.t. The solid precipitate was isolated and digirated with 2-propanol, dried in vacuum, dissolved in DCM and washed with citric acid. 4-{[4-(methoxycarbonyl)-phenyl]amino}-4-oxo-3-phenylbutanoic acid was isolated as a crystalline solid (980 mg, 2.88 mmol). M.p. 212°C; TLC (cyclohexane/EtOAc 7/3) Rf 0.17; 1H NMR (400 MHz, D6-DMSO) δ 1.39 (t, J=7.2 Hz, 3H); 2.75 (dd, J=3.7 Hz, J=17.2 Hz, 1H); 3.42 (dd, J=3.7 Hz, J=17.2 Hz, 1H); 4.09 (dd, J=3.7 Hz, J=17.2 Hz, 1H); 4.35 (q, J=7.2 Hz, 2H); 7.33-7.41 (m, 5H), 7.75 (mc, 2H), 7.94 (mc, 2H). The regiochemistry was determined using NOESY NMR spectroscopy.
    • Example V: 4-{[4-(ethoxycarbonyl)phenyl]amino}-4-oxo-3-phenylbutanoic acid
  • Figure imgb0011
  • The solvent was evaporated and the residue was purified by flash chromatography yielding 4-{[4-(methoxycarbonyl)phenyl]amino}-4-oxo-3-phenylbutanoic acid (690 mg, 2.87 mmol) M.p. 190°C; TLC (DCM/MeOH 9/1) Rf 0.21; 1H NMR (400 MHz, D6-DMSO) δ 1.30 (t, J=7.1 Hz, 3H); 2.23 (s, 3H); 2.80 (mc, 1H); 3.25 (mc, 1H); 4.25-4.30 (m, 3H); 3.74 (s, 3H); 6.00 (bs, 1H); 6.65 (d, J=6.6 Hz, 2H); 6.90-6.96 (m, 2H); 7.10-7.19 (m, 4H); 7.34-7.38 (m, 2H); 7.43-7.51 (m, 2H); 7.69-7.71 (m, 1H); 7.79-7.90 (m, 2H); 8.70 (s, 1H); 8.97 (s, 1H).
    • Step B Example V: Ethyl 4-[(4-{[4-({[(2-methylphenyl)amino]carbonyl}amino)phenyl]-amino}-4-oxo-3-phenylbutanoyl)amino]benzoate
  • Figure imgb0012
  • 4-{[4-(Ethoxycarbonyl)phenyl]amino}-4-oxo-2-phenylbutanoic acid (400 mg, 1.17 mmol) was dissolved in MeCN (10 mL), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimidhydrochlorid (247 mg, 1.29 mmol), 1-hydroxy-1H-benzotriazol (158 mg, 1.17 mmol) followed by N-(4-aminophenyl)-N'-(2-methylphenyl)urea (282 mg, 1.17 mmol) was added at r.t and the reaction mixture was stirred for 24 h. Ethyl 4-[(4-{[4-({[(2-methylphenyl)amino]carbonyl}amino)phenyl]amino}-4-oxo-3-phenyl-butanoyl)amino]benzoate (194mg, 0.34 mmol) was isolated by filtration as a colorless solid. M.p. >220°C; TLC (DCM/MeOH 9/1) Rf 0.21; 1H NMR (400 MHz, D6-DMSO) δ 1.30 (t, J=7.1 Hz, 3H); 2.23 (s, 3H); 2.80 (mc, 1H); 3.25 (mc, 1H); 4.25-4.30 (m, 3H); 3.74 (s, 3H); 6.00 (bs, 1H); 6.65 (d, J=6.6 Hz, 2H); 6.90-6.96 (m, 2H); 7.10-7.19 (m, 4H); 7.34-7.38 (m, 2H); 7.43-7.51 (m, 2H); 7.69-7.71 (m, 1H); 7.79-7.90 (m, 2H); 8.70 (s, 1H); 8.97 (s, 1H).
    Figure imgb0013
    • Step C Example 1: 4-[(4-{[4-({[(2-Methylphenyl)amino]carbonyl}amino)phenyl]-amino}-4-oxo-3-phenylbutanoyl)amino]benzoic acid
  • Figure imgb0014
  • Ethyl 4-[(4-{[4-({[(2-methylphenyl)amino]carbonyl}amino)phenyl]amino}-4-oxo-3-phenylbutanoyl)amino]benzoate (160 mg, 0.28 mmol) was dissolved in water:THF (10 mL; 1:1;v:v) and LiOH (27 mg, 1.13 mmol) was added at 0°C and the reaction mixture was stirred at r.t. for 24h. The reaction mixture was acidified, the product was isolated by filtration and purified by flash chromatography (CH2Cl2/MeOH/AcOH; 10/1/0.5) (11 mg, 0.02 mmol). M.p. 169°C. TLC (DCM/MeOH 9/1) Rf 0.27, ESI-MS: 537[M+H]+.
    Figure imgb0015
    • In vitro assay: adhesion of Ramos cells to immobilized VCAM-1 (domains 1-3) Preparation of VCAM-1 (extracellular domains 1-3)
  • Complementary DNA (cDNA) encoding 7-domain form of VCAM-1 (GenBank accession #M60335) was obtained using Rapid-ScreenTM cDNA library panels (OriGene Technologies, Inc) at Takara Gene Analysis Center (Shiga, Japan). The primers used were 5'-CCA AGG CAG AGT ACG CAA AC-3' (sense) and 5'-TGG CAG GTA TTA TTA AGG AG-3' (antisense). PCR amplification of the 3-domain VCAM-1 cDNA was perform using Pfu DNA polymerase (Stratagene) with the following sets of primers: (U-VCAMd1-3) 5'-CCA TAT GGT ACC TGA TCA ATT TAA AAT CGA GAC CAC CCC AGA A-3'; (L-VCAMd1-3) 5'-CCA TAT AGC AAT CCT AGG TCC AGG GGA GAT CTC AAC AGT AAA-3'. PCR cycle was 94°C for 45 sec, 55°C for 45 sec, 72°C for 2 min, repeating 15 cycles. After the purification of the PCR product, the fragment was digested with KpnI-AvrII. The digested fragment was ligated into pBluescript IISK(-) (Strategene), which was linearized by digesting with KpnI-XhoI. The ligation was followed by transformation to a Dam/Dcm methylase-free E. coli strain SCS110 (Strategene) to create the donor plasmid pHH7. To direct VCAM-1 molecule into the insect cell secretory pathway, the VCAM-1 coding sequence was fused to signal peptide sequence of honeybee melittin. The resulting melittin-VCAM fusion was placed in correct orientation to the baculovirus polyhedrin promoter. Baculovirus transfer vector containing first 3-domain form VCAM-1 (pH10) was constructed by ligation of 0.9 kb fragment from AvrII/Klenow/BclI digests of pH7 into SalI/Klenow/BamHI digests of pMelBacB (Invitrogen). Recombinant baculovirus was generated by using Bac-N-Blue™ Transfection kit (Invitrogen) according to the manufacture's instruction. The recombinant virus was amplified by infection to High-Five™ insect cells for 5 - 6 days, and virus titer was determined by plaque assay.
  • High-Five™ insect cells were pelleted in a 225 ml conical tube by centrifugation at 1000 rpm for 5 min. After discarding the supernatant, the pellet was resuspended in 1.5 x 109 pfu (MOI = 5) of high-titer virus solution, followed by incubation for 1.5 hours at room temperature. The cells were pelleted again and washed once in fresh Express Five™ serum free medium. The cells were pelleted again and finally, resuspended in 200 ml of fresh Express Five TM medium, transferred to a 1,000 ml shaker flask, and incubated in a shaker at 27°C, 130 rpm, for 48 hours before the culture supernatant was collected. The purification of 3-domain form of VCAM-1 from the culture supernatant was performed by one-step anion exchange chromatography. Protein concentration was determined by using Coomassie protein assay reagent (Pierce) according to the manufacture's instruction.
    • Preparation of VCAM-1 coated microtiter plates
  • Recombinant human VCAM-1 (extracellular domains 1-3) was dissolved at 1.0 µg/ml in PBS. Each well of the microtiter plates (Nalge Nunc International, Fluoronunc Cert, 437958) was coated with 100 µl of substrate or for background control with buffer alone for 15 hours at 4°C. After discarding the substrate solution, the wells were blocked using 150 µl per well of block solution (Kirkegaard Perry Laboratories, 50-61-01) for 90 minutes. The plate was washed with wash buffer containing 24 mM Tris-HCl (pH 7.4), 137 mM NaCl, 27 mM KCl and 2 mM MnCl2 just before addition of the assay.
    • In Vitro Assay using Ramos cells Preparation of fluorescence labeled Ramos cells
  • Ramos cells (American Type Culture Collection, Clone CRL-1596) were cultured in RPMI 1640 medium (Nikken Bio Medical Laboratory, CM1101) supplemented with 10% fetal bovine serum (Hyclone, A-1119-L), 100 U/ml penicilin (Gibco BRL, 15140-122) and 100 µg/ml streptomycin (Gibco BRL, 15140-122) in a humidified incubator at 37°C with 5% CO2.
  • Ramos cells were incubated with phosphate balanced solution (PBS, Nissui, 05913) containing 25 µM of 5(-and -6)-carboxyfluorescein diacetate, succinimidyle ester (CFSE, Dojindo Laboratories, 345-06441) for 20 min at room temperature while gently swirling every 5 min. After centrifugation at 1000 rpm for 5 min, the cell pellet was resuspended with adhesion assay buffer at a cell density of 4 x 106 cells/ml. The adhesion assay buffer was composed of 24 mM Tris-HCl (pH 7.4), 137 mM NaCl, 27 mM KCl, 4 mM glucose, 0.1 % bovine serum albumin (BSA, Sigma, A9647) and 2 mM MnCl2.
    • Assay procedure (Ramos cells)
  • The assay solution containing each test compounds or 5 µg/ml anti-CD49d monoclonal antibody (Immunotech, 0764) was transferred to the VCAM-1 coated plates. The final concentration of each test compounds was 5 µM, 10 µM or various concentrations ranging from 0.0001 µM to 10 µM using a standard 5-point serial dilution. The assay solution containing the labeled Ramos cells was transferred to the VCAM-1 coated plates at a cell density of 2 x 105 cells per well and incubated for 1 hour at 37°C. The non-adherent cells were removed by washing the plates 3 times with wash buffer. The adherent cells were broken by addition of 1 % Triton X-100 (Nacalai Tesque, 355-01). Released CFSC was quantified fluorescence measurement in a fluorometer (Wallac, ARVO 1420 multilabel counter).
  • The adhesion of Ramos cells to VCAM-1 was analyzed by percent binding calculated by the formula:
  • 100 x (FTS - FBG) / (FTB - FBG) = % binding, where FTB is the total fluorescent intensity from VCAM-1 coated wells without test compound; FBG is the fluorescent intensity from wells with anti-CD49d monoclonal antibody and FTS is the fluorescent intensity from wells containing the test compound of this invention.
    • In vitro activity
  • In the Ramos VCAM-1 assay the observed IC50 value ranges are indicated in Table 4.
    C > 10 µM ≥ B > 1 µM ≥ A Table 4
    No IC50
    1 A
    2 C
    3 C
    4 A

Claims (12)

  1. Compounds of the general formula (I),
    Figure imgb0016
    wherein
    R1 represents a 4- to 9-membered saturated, unsaturated or aromatic cyclic residue,
    which can contain 0 to 3 heteroatoms selected independently from the group N, S and O,
    and wherein R1 is substituted by -R1-1-Z,
    wherein
    R1-1 represents a bond, -O-, -S-, NR1-2, C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C6 or C10 aryl, C3-C7 cycloalkyl or a 4-9-membered saturated or unsaturated heterocyclic residue containing up to 3 heteroatoms selected from the group oxygen, nitrogen or sulfur,
    wherein R1-1 can optionally be substituted by 1 to 2 substituents selected from the group R1-3,
    wherein R1-2 can optionally be hydrogen, C1-C10 alkyl, C2-C10 alkenyl or C2-C10 alkynyl, and
    wherein R1-3 represents hydrogen, C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C6 or C10 aryl, C3-C7 cycloalkyl or a 4-9-membered saturated or unsaturated heterocyclic residue containing up to 3 heteroatoms selected from the group oxygen, nitrogen or sulfur,
    Z represents -C(O)ORZ-1, -C(O)NRZ-2RZ-3, -SO2NRZ-2RZ-3, -SO(ORZ-1), -SO2(ORZ-1), -P(O)RZ-1(ORZ-3) or -PO(ORZ-1)(ORZ-3),
    wherein RZ-2 is hydrogen, C1-C4 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C6 or C10 aryl, -C(O)RZ-4 or -SO2RZ-4,
    wherein RZ-4 is C1-C4 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C6 or C10 aryl,
    RZ-1 and RZ-3 are independently selected from the group hydrogen, C1-C4 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C6 or C10 aryl or benzyl,
    wherein RZ-1 and RZ-3 can optionally be substituted by 1 to 3 substituents selected from the group C1-C4 alkyl, C1-C4 alkyloxy, halogen, nitro, cyano,

    and wherein R1 can optionally be substituted by 0 to 2 substituents R1-4, halogen, nitro, amino, cyano and oxo,
    wherein
    R1-4 is selected from the group C1-C4 alkyl, C1-C4 alkyloxy, phenyl, phenoxy, phenylamino, C3-C6 cycloalkyl,
    R2 represents hydrogen, C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C6 or C10 aryl or C3-C7 cycloalkyl,

    wherein R2 can optionally be substituted by 1 to 3 radicals independently selected from the group C1-C4 alkyl, trifluormethyl, trifluormethoxy, halogen, cyano, nitro or oxo,
    R3 represents hydrogen, C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C6 or C10 aryl, C3-C7 cycloalkyl or a 4-9-membered saturated or unsaturated heterocyclic residue containing up to 2 heteroatoms selected from the group oxygen, nitrogen or sulfur,

    wherein R3 can optionally be substituted by 1 to 3 radicals R3-1,
    wherein R3-1 represents C1-C4 alkyl, trifluormethyl, trifluormethoxy, -OR3-2, -SR3-2, NR3-3R3-4, -C(O)R3-2, S(O)R3-2, -SO2R3-2, -OC(O)R3-2, -C(O)NR3-3R3-4, -NR3-2C(O)R3-3, -SO2NR3-3R3-4, NR3-2SO2R3-3, -NR3-2C(O)NR3-3R3-4, -NR3-2C(O)OR3-3, -OC(O)NR3-3R3-4, -CO2R3-5, halogen, cyano, nitro or oxo,
    wherein R3-2 represents hydrogen, C1-C4 alkyl, C3-C6 cycloalkyl, C6 or C10 aryl,
    wherein R3-3 and R3-4 are independently selected from the group hydrogen, C1-C4 alkyl, C3-C6 cycloalkyl, C6 or C10 aryl or benzyl,
    and wherein R3-5 represents C1-C4 alkyl, C3-C6 cycloalkyl, C6 or C10 aryl,
    R4 represents hydrogen, C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C6 or C10 aryl, C3-C7 cycloalkyl or a 4-9-membered saturated or unsatu- rated heterocyclic residue containing up to 2 heteroatoms selected from the group oxygen, nitrogen or sulfur,

    wherein R4 can optionally be substituted by 1 to 3 radicals R4-1,
    wherein R4-1 represents C1-C4 alkyl, trifluormethyl, trifluormethoxy, -OR4-2, -SR4-2, NR4-3R4-4, -C(O)R4-2, S(O)R4-2, -SO2R4-2, -OC(O)R4-2, -C(O)NR4-R4-4, -NR4-2C(O)R4-3, -SO2NR4-3R4-4, NR4-2SO2R4-3, -NR4-2C(O)NR4-3R4-4, -NR4-2C(O)OR4-3, -OC(O)NR4-3R4-4, -CO2R4-5, halogen, cyano, nitro or oxo,
    wherein R4-2 represents hydrogen, C1-C4 alkyl, C3-C6 cycloalkyl, C6 or C10 aryl,
    wherein R4-3 and R4-4 are independently selected from the group hydrogen, C1-C4 alkyl, C3-C6 cycloalkyl, C6 or C10 aryl or benzyl,
    and wherein R4-5 represents C1-C4 alkyl, C3-C6 cycloalkyl, C6 or C10 aryl
    or
    R3 and R4 together with the carbon atom to which they are attached form a 4-7-membered saturated or unsaturated ring containing up to 2 heteroatoms selected from the group oxygen, nitrogen or sulfur, which can optionally be substituted by 1 to 2 substituents selected from the group C1-C4 alkyl, phenyl, benzyl, C3-C7 cycloalkyl, C1-C4 alkyloxy, halogen, nitro, cyano, oxo and which can be fused with a 3-7 membered homocyclic or heterocyclic, saturated, unsaturated or aromatic ring,
    R5 represents hydrogen, C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C6 or C10 aryl or C3-C7 cycloalkyl,
    wherein R5 can optionally up to threefoldedly be substituted by C1-C4 alkyl, trifluormethyl, trifluormethoxy, halogen, cyano, nitro or oxo,
    R6 represents phenyl or a 5- to 6-membered aromatic heterocyclic residue containing up to 3 heteroatoms independently selected from the group oxygen, nitrogen or sulfur,
    wherein R6 is substituted by -NR6-2C(O)NR6-3R6-4 or -NR6-2C(S)NR6-3R6-4 and can furthermore optionally be substituted by halogen,
    wherein R6-2 and R6-3 are independently selected from the group hydrogen or C1-C4 alkyl, or together form a group
    Figure imgb0017
    and wherein R6-4 represents phenyl,
    wherein R6-4 can optionally be substituted by 1-2 substituents selected from the group C1-C4 alkyl, C1-C4 alkyloxy, halogen, nitro, trifluoromethyl, trifluoromethoxy or cyano,
    or
    R6 represents a group
    Figure imgb0018
    wherein R6-1 represents a substituent selected from the group hydrogen, C1-C4 alkyl, C1-C4 alkyloxy, halogen, nitro, trifluoromethyl, trifluoromethoxy or cyano,
    and
    wherein R6-5 represents a substituent selected from the group hydrogen, C1-C4 alkyl, C1-C4 alkyloxy, halogen, nitro, trifluoromethyl, trifluoromethoxy or cyano,
    R7 represents hydrogen, C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C6 or C10 aryl, C3-C7 cycloalkyl or a 4-9-membered saturated or unsaturated heterocyclic residue containing up to 2 heteroatoms selected from the group oxygen, nitrogen or sulfur,

    wherein R7 can optionally be substituted by 1 to 3 radicals R7-1,
    wherein R7-1 represents C1-C4 alkyl, trifluormethyl, trifluormethoxy, -OR7-2, -SR7-2, NR7-3R7-4, -C(O)R7-2, S(O)R7-2, -SO2R7-2, -OC(O)R7-2, -C(O)NR7-3R7-4, -NR7-2C(O)R7-3, -SO2NR7-3R7-4, NR7-2SO2R7-3, -NR7-2C(O)NR7-3R7-4, -NR7-2C(O)OR7-3, -OC(O)NR7-3R7-4, -CO2R7-5, halogen, cyano, nitro or oxo,
    wherein R7-2 represents hydrogen, C1-C4 alkyl, C3-C6 cycloalkyl, C6 or C10 aryl,
    wherein R7-3 and R7-4 are independently selected from the group hydrogen, C1-C4 alkyl, C3-C6 cycloalkyl, C6 or C10 aryl or benzyl,
    and wherein R7-5 represents C1-C4 alkyl, C3-C6 cycloalkyl, C6 or C10 aryl
    R8 represents hydrogen, C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C6 or C10 aryl, C3-C7 cycloalkyl or a 4-9-membered saturated or unsaturated heterocyclic residue containing up to 2 heteroatoms selected from the group oxygen, nitrogen or sulfur,

    wherein R8 can optionally be substituted by 1 to 3 radicals R8-1,
    wherein R8-1 represents C1-C4 alkyl, trifluormethyl, trifluormethoxy, -OR8-2, -SR8-2, NR8-3R8-4, -C(O)R8-2, S(O)R8-2, -SO2R8-2, -OC(O)R8-2, -C(O)NR8-3R8-4, -NR8-2C(O)R8-3, -SO2NR8-3R8-4, NR8-2SO2R8-3, -NR8-2C(O)NR8-3R8-4, -NR8-2C(O)OR8-3, -OC(O)NR8-3R8-4, -CO2R8-5, halogen, cyano, nitro or oxo,
    wherein R8-2 represents hydrogen, C1-C4 alkyl, C3-C6 cycloalkyl, C6 or C10 aryl,
    wherein R8-3 and R8-4 are independently selected from the group hydrogen, C1-C4 alkyl, C3-C6 cycloalkyl, C6 or C10 aryl or benzyl,
    and wherein R8-5 represents C1-C4 alkyl, C3-C6 cycloalkyl, C6 or C10 aryl
    or
    R7 and R8 together form a 4-7-membered saturated or unsaturated ring containing up to 2 heteroatoms selected from the group oxygen, nitrogen or sulfur, which can optionally be substituted by 1 to 2 substituents selected from the group C1-C4 alkyl, phenyl, benzyl, C3-C7 cycloalkyl, C1-C4 alkyloxy, halogen, nitro, cyano, oxo and which can be fused with a 3-7 membered homocyclic or heterocyclic, saturated, unsaturated or aromatic ring,
    X represents bond or (-CRX-1RX-2-)n,

    wherein RX-1 and RX-2 can be independently selected from the group hydrogen, C1-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl,
    wherein RX-1 and RX-2 can optionally independently be substituted by 1 to 2 substituents selected from the group C1-C4 alkyl, phenyl, benzyl, C3-C7 cycloalkyl, C1-C4 alkyloxy, halogen, nitro, cyano, oxo,
    and wherein n is an integer 0 or 1,
    and pharmaceutically acceptable salts thereof.
  2. Compounds of general formula (I) according to claim 1,
    wherein R1 represents a phenyl ring.
  3. Compounds according to claim 1 or 2,
    wherein R1-1 represents a bond and Z represents COORZ-1,
    wherein RZ-1 has the meaning indicated above.
  4. Compounds according to claim 1, 2 or 3,
    wherein R6 represents phenyl, which is substituted by -NHC(O)NHR6-4,
    wherein R6-4 is substituted with methyl or trifluoromethoxy.
  5. Compounds according to any one of claims 1 to 4,
    wherein X represents a methylene group.
  6. Compounds according to any one of claims 1 to 5,
    wherein one of the substituents R3, R4, R7 and R8 represents phenyl, which can optionally be substituted by up to three substituents independently selected from the group C1-C4-alkyl and halogen, and the remaining substituents represent hydrogen.
  7. A process for preparation of compounds of general formula (I) according to any one of claims 1 to 6, which comprises reaction of carboxylic acids of general formula (I')
    Figure imgb0019
    or activated derivatives thereof,
    wherein
    R1, R2, R3, R4, R7 and R8 have the above-mentioned meaning,
    with compounds of the general formula (I")

            R6-X-NR5H     (I")

    wherein
    X, R5 and R6 have the above-mentioned meaning,
    in inert solvents.
  8. Compounds according to any one of claims 1 to 6 for the treatment of diseases.
  9. The use of a compound according to any one of claims 1 to 6 in the manufacture of a medicament for the treatment or the prevention of a condition mediated by integrins.
  10. The use of a compound according to any one of claims 1 to 6 in the manufacture of a medicament for the treatment or the prevention of atherosclerosis, asthma, chronic obstructive pulmonary disease (COPD), allergies, diabetes, inflammatory bowel disease, multiple sclerosis, myocardial ischemia, rheumatoid arthritis, transplant rejection and other inflammatory, autoimmune and immune disorders.
  11. Pharmaceutical composition, comprising compounds according to any one of claims 1 to 6 and a pharmaceutically acceptable carrier.
  12. Compounds of general formula (I) according to claim 2, characterized in that R1 is a 1,4-substituted phenyl ring.
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JP4617449B2 (en) 2002-07-11 2011-01-26 ヴィキュロン ファーマシューティカルズ インコーポレイテッド N-hydroxyamide derivative having antibacterial activity
CA2770493A1 (en) 2003-07-24 2005-02-03 Daiichi Pharmaceutical Co., Ltd. Cyclohexanecarboxylic acid compound
SG176628A1 (en) 2009-06-05 2012-01-30 Link Medicine Corp Aminopyrrolidinone derivatives and uses thereof

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DE10006453A1 (en) 2000-02-14 2001-08-16 Bayer Ag New piperidylcarboxylic acid derivatives, are integrin antagonists useful for treating inflammatory, autoimmune or immunological diseases, e.g. atherosclerosis, asthma, diabetes, rheumatoid arthritis or transplant rejection
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GB2369357A (en) 2000-10-09 2002-05-29 Bayer Ag Aliphatic, cyclic amino carboxylic acids as integrin antagonists
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GB2367816A (en) * 2000-10-09 2002-04-17 Bayer Ag Urea- and thiourea-containing derivatives of beta-amino acids

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US20040254157A1 (en) 2004-12-16
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EP1406865A2 (en) 2004-04-14
GB0116577D0 (en) 2001-08-29
PT1406865E (en) 2006-06-30
US7064229B2 (en) 2006-06-20
DE60209122T2 (en) 2006-08-03
ATE317380T1 (en) 2006-02-15
ES2258636T3 (en) 2006-09-01
DK1406865T3 (en) 2006-05-08
GB2377933A (en) 2003-01-29

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